Iterative, multiplexed CRISPR-mediated gene editing for functional analysis of complex protease gene clusters.

ABSTRACT: Elucidation of gene function by reverse genetics in animal models frequently is complicated by the functional redundancy of homologous genes. This obstacle often is compounded by the tight clustering of homologous genes, which precludes the generation of multigene-deficient animals through standard interbreeding of single-deficient animals. Here, we describe an iterative, multiplexed CRISPR-based approach for simultaneous gene editing in the complex seven-member human airway trypsin-like protease/differentially expressed in a squamous cell carcinoma (HAT/DESC) cluster of membrane-anchored serine proteases. Through four cycles of targeting, we generated a library of 18 unique congenic mouse strains lacking combinations of HAT/DESC proteases, including a mouse strain deficient in all seven p

SUBMITTER: Callies LK 

PROVIDER: S-EPMC6827298 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature